Recombinant arthrobotrys oligospora of Aozl gene dual promoters and preparation method of recombinant arthrobotrys oligospora

Abstract

The invention discloses recombinant arthrobotrys oligospora of Aozl gene dual promoters, which is used for preventing and controlling ruminant animal digestive tract nematode diseases caused by nemathelminthes nematode, and a preparation method of the recombinant arthrobotrys oligospora. The Aozl gene promoters of arthrobotrys oligospora are cloned by adopting a chromosome DNA (Deoxyribonucleic Acid) walking technology, and the recombinant arthrobotrys oligospora efficiently expressing the Aozl gene dual promoters is constructed by utilizing an agrobacterium tumefaciens-mediated fungus transformation technology. Compared with a traditional genetic transformation method, the method provided by the invention has the advantages of wide recipient material range, high transformation efficiency,stable inheritance transformant and the like, and the bottleneck of a traditional transformation method is overcome. Compared with a non-recombinant maternal arthrobotrys oligospora strain, the strain obtained by the method has good genetic stability; the Aozl gene transcriptional level is enhanced and the nematode-trapping activity is improved; an efficient gene engineering strain is provided for researches and development of a biological prevention and control preparation for the ruminant animal digestive tract nematode diseases.

Description

technical field [0001] The invention relates to Arthrobuspora oligospora serine protease Aoz1 gene dual-promoter recombinant Arthrobuspora oligospora and a preparation method thereof, belonging to the field of biotechnology. technical background [0002] Ruminant digestive nematode disease is a kind of parasitic disease caused by various nematodes belonging to the nematode class Nematoda parasitizing in the digestive tract of ruminants, which seriously endangers the sustainable and healthy development of my country's aquaculture industry. According to statistics, among ruminant helminth diseases, digestive tract nematode infection accounts for more than half, among which Trichostrongylus, Haemonchus, Leptospirosis, Oster, Marshall, and Trichocerus Mainly. In recent years, with the continuous strengthening of the country's prevention and control measures for animal infectious diseases, the infectious diseases of ruminants have been effectively controlled, and the hazards of ...

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Problems solved by technology

However, although Arthrobuspora oligospora isolated from nature showed strong nematode-predating activity at the initial stage of isolation, with the increase in the number of subcultures in the medium, the expression level of extracellular proteolytic enzymes also decreased, and the nematode-predating activity will gradually decrease, which has become a bottleneck restricting the development of efficient nematode-predating biological agents

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